Anthelmintic 5-arylazo rhodanines



United States Patent Oflice Patented July 16, 1953 3,098,007 ANI'HELMINTIC S-ARYLAZO RHODANINES Gerald Brody, Quincy, lll., assignor to Moorman Manufacturing Company, Quincy, 111., a corporation of Illinois No Drawing. Filed Jan. 17, 1062, Ser. No. 166,905

13 Claims. (Cl. 16753.1)

in which R represents the hydrogen, methyl, ethyl, propyl, or butyl groups and X represents methyl, chloro and nitro substituents.

The large poultry roundworm, A. gallf, is extremely common and often results in decreased feed intake, retarded growth, increased susceptibility to other disease organisms, and if present in sufficient numbers, in death of the bird. As many as 600 worms averaging three inches in length have been recovered from a naturally infected bird.

This parasitic infection is spread by mature female worms which produce worm eggs in great numbers which are passed out into the external environment with chicken fecal droppings. Under ideal environmental conditions of moisture and temperature, the eggs mature and can become infective in about two weeks. Ingestion of infective eggs by other birds completes the life cycle, and in about fifty days the newly ingested infective eggs produce mature worms capable of reproduction.

Approximately one-third of all dogs harbor internal worms with ascarids and hookworms constituting the commonest types of worm infection in dogs. In general, ascarids and hookworms in dogs can cause nutritional deficiencies resulting in depressed growth and development. Dog hookworms are blood suckers and are especially dangerous to young pups causing anemia, unthriftiness and even death of entire litters of pups. Toxocara canis, the commonest type of ascarid in dogs, has a complex life cycle with the larvae migrating to various parts of the body, especially the liver and lungs, before maturation in the small intestine. This migratory phase can cause severe pneumonia in growing pups, and in pregnant dogs can cause prenatal infection of newly born pups contributing to weak or still-born pups.

The active ingredients of this invention effectively expel mature A. galli worms when administered in poultry feed at 0.00625%0.20% by weight in the ration for one to four days. When administered continuously at 0.0125- O.10% by weight in the ration, starting at the time of exposure to infective ascarid eggs, these compounds are highly effective in expelling immature worms and retarding the growth and development of those worms which remain in the bird. Thus, these compounds administered continuously in feed to young birds can break the life cycle of this important poultry parasite.

The active ingredients of this invention also effectively expel mature dog ascarids and hookworms when administered in the ration of the dog. Although the optimal dosage for a given dog varies with the age and weight of the dog, in general, good results were obtained when the total drug intake ranged from 1004.50 rug/kg. body weight administered over a 1-5 day treatment period. When administered at lower levels (4.4-8.9 mg./kg./ day) for longer periods of time, these compounds suppress the production of ascarid and hookwork eggs. The percent age of drugs in the feed would vary with the age and body weight of the dog. For the 1-5 day treatment the percentage of drug in feed varied from 0.022-0.33% and as low as 0.0088% for the low level feeding program.

Accordingly, the object of this invention, generally stated, is the provision of new and improved medicated feed formulations, compositions and rations which are safe, reliable, efiicient and economical when used for the elimination, treatment, control and prevention of the large poultry roundworrns, A. galli, in poultry and ascarid and hookwork infection in dogs.

Certain other objects of the invention will in part be obvious and will in part appear hereinafter.

For a more complete understanding of the nature and scope of the present invention, reference may be made to the following detailed description and illustrative examples which may be used to practice the invention and which will suggest other and additional embodiments of the invention to those skilled in the art.

EXAMPLE 1 Mature birds (chickens) which had been exposed to natural worm infections were housed in individual cages and fecal samples taken to ascertain presence of A. galii infections. All birds shown to harbor this parasite were subsequently used in anthelmintic tests. Medicated feeds containing these 5(n itrophenylazo) rhodanine compounds were prepared by admixing 1 gram of active ingredient in 1000 grams poultry ration (2 parts of ground corn and 1 part of commercial chick concentrate) to produce a medicated feed containing 0.10% active ingredient by weight. In a similar manner, medicated feeds containing other concentrations of active ingredient were prepared.

In experiments 108 and 131, the birds were weighed, housed in individual cages and put on medicated feed containing 0.10% B-methy'l, 5(P-nitrophenylazo) rhodanine for 7 and 3 days, respectively. All droppings eliminated from the test birds were carefully examined for expelled worms during the entire period of medication. The day following termination of medication, the birds were sacrificed and all worms remaining in the intestinal tract were recorded. The initial worm infection represents the sum of the worms expelled and those remaining in the gut. Efiicacy of treatment is calculated as follows:

Number of worms expelled Total initial infection X Efiicacy (percent) In experiment 110, naturally infected birds were allotted at random into two groups of seven birds per lot. One group was fed 0.10% S-methyl, 5(P-nitrophenylazo) rhodanine for seven days whereas the other group did not receive any medication and thus served as non-medicated control. At the end of the seven-day experimental period, all birds in botlrlots were sacrificed and the number of worms found in the intestinal tracts were recorded. Efficacy of treatment was ascertained by comparison of the worm burdens present in the treated birds as compared to the non-treated birds.

The results of these three experiments, which are summarized in Table 1, clearly indicate the excellent anthelmintic activity of 3-methyl, 5(P-nitrophenylazo) rhodanine.

Table 1 ANTIIELMINTIC ACTIVITY OF 3-METHYL, 5(P-NITROPHENYLAZO) RHODANINE ADMINISTERED AT 0.10% IN POULTRY FEED AGAINST MATURE ASCARIDIA GALLI INFECTIONS Number Total drug Total worms Expt. days on intake NuIn- Eilieaoy Feeding method Number medicaper bird her (percent) tion (mg/kg.) birds Initial At autopsy Individual cages -108 7 425. 6 8 38 0 100. 0 (3-131 3 159. 6 4 60 2 96. 7

Grouped blrds (3-110 0 0 7 154 G-110 7 590. 9 7 6 96.1

EXAMPLE 2 15 As seen in Table 2, administration of the anthelmintics h g anthelmmtlc dactwlty of Ether g lj a g at levels of 0.00625 %0.10% in feed for 1-4 days was r o anine compoun s using tee niques escri e in t e 7 t previous example was studied and the following comvery efiecnve m expenmg ma ure poultry ascands' Table 2 ANTHELMINTIG ACTIVITY OF -ARYLAZO RHODANINES ADMINISTEREI) AT SEVERAL LEVELS IN FEED FOR SEVERAL INTERVALS OF TIME AGAINST MATURE ASCARIDIA GALLI INFECTIONS Substituent on rhodauinc ring Total Total Worms Percent Number drug Num- Feedlng method (Expt. No.) drug days on intake bur Efficacy in feed mediper bird birds At (percent) cation (l-position fi-positiou (lringjl Initial autopsy Individual cages (Ci-139 and G446)-.. 0.10 3 M ethyl. P-nltropheuylezo 187.6 1 1 0 100.0 Ethyl .d0 -7 169. 7 2 13 0 100, O n-Propy1 .do- 182. 5 2 401 0 100.0 Hydrogen- .do. 185. 3 3 82 1 98.8 0 2 d0 d0-.- 101.3 5 207 4 98.5 0 05 3 Methyl.-- do.. 62.2 1 6 0 100.0 Ethyl 0..- 75. 2 3 681 22 96.8 n-Propyl ..do 91. 6 3 414 85 79.4 Hydrogeno 72.3 5 753 204 72,9 Methyl.-- 2,4 dinitrophenylazo- 54.2 2 134 85.8 0.05 2 Hydrogen. P-nitrophonylazo 58. 7 5 731 288 60. 6 0.20 1 do 0 90. 3 5 258 90,3

Grouped birds ((3-211) 0. 026 4 Methyl.-. .do 96. 5 8 97. 7 n-Buty do 102.2 8 98.7 Methyl 2,4 dinitrophenylazmnn 102.0 8 72.6 d0 2 methyl, 4 nitrophenylazo. S6. 0 8 100. 0 do 2 chloro, 4 nitrophonylazo... 100.3 8 100.0 N omrnedicated control 0 8 Grouped birds (0-213) 0.0075 4 Methyl. 2 methyl, 4 nltrophenylazd. 24. 8 8 4 99. 7 do.-. 2 chloro, 4 nitrophenylazo.-. 32.1 8 G9 95. 4 Nonmedicated control 0 7 1. 301

Individual cages (Ci-212B) 0. 00625 4 Methyl.-. 2 methyl, 4 nitrophenylozo. 21 9 6 1,096 31 97, 2 do 2 chloro, 4 nitrophenylazo 22 2 6 5 7 99. 2

pounds were found to exhibit good anthelmmtlc activity: A summary f anthelmmtlc actlvlty f the vanous 5- 5(P-nitrophenylazo) rhodanine arylazo rhodarunes against mature poultry ascarids 1s 3-methyl, 5(P-n1trophenylazo) rhodanine 59 presented in Table 3.

Table 3 SUMMARY OF ANTHELMINTIC ACTIVITY OF 5 ARYLAZO RHODANINES ADIVIINIS TE RED IN FEED AGAINST MATURE POULTRY ASCARIDSINDIVIDUAL CAGED BIRD ST UDIES Percent anthelmintie activity at various levels of total Substituont on rhodanine ring drug; intake expressed as mg./kg.; equals number of 3-posltion 5-1Jositi0n 100 75 50 12. 5

Hydrogen 4nitrophcnylazo 83.6 (11) 81.6 7) 73.5 4) 2.3 2) Methyldo 97.3 (20) 94.7 (12) 77.9 07.6 (24) Ethyl-.- 100.0 4 95.2 s 66.7 5 73.0 (10) nPropyl 9243 0) 100.0 5) 37.7 (16) 04.9 7 i-Proply 97.2 (3) 82.0 (o) 87.5 5) 07.0 (13) n-But 1 100.0 (3) 100.0 6) sea (18) 82.1 (3) Methyl 2,4 dinitrophenylam 75.4 8) 58.8 1) Do Z-ehloro, 4 nltrophenylazo. 100. 0 2) 99. 2 8) Do Z-methyl, 4 nitrophenylazo 100.0 2) 97.7 9i

3-ethyl, 5(P-nitrophenylazo) rhodanine EXAMPLE 3 -P PYL P Y fhodamfle In order to evaluate activity of these compounds against P PYL p y l'hodafllne immature stages of A. galli, one-Week-old White Leghorn yl, qpl y rhodamne chicks were artificially infected with 300-550 infective 31 p l y Thodafilfle Worm eggs per bird. Medicated feeds containing an active 3-methyl, 5(2'rnethyl, 4n1tr0phenylazo) rhodanlne drug concentration ranging from 0.025-0.10% by weight 3-methyl, 5(2chloro, 4'nitrophenylazo) rhodanine were administered continuously from the time of infection when the birds were one week of age until the infection was four weeks old at which time all birds were sacrificed and worm burdens of the various experimental groups ascertained. Typical results, presented in Table 4 below indicate that continuous feeding of 3-methyl, 5(P-nitrophenylazo) rhodanine, 5(P-nitrophenylazo) rhodanine, B-methyl, 5(2' methyl, 4' nitrophenylazo) rhoctanine, and S-rnethyl, 5(2 chloro, 4 nitrophenylazo) rhodanine is highly elTective in controlling immature A. galli infections. Similar tests have demonstrated comparable activity of the other members of this series against immature A. galli infections.

Table 4 EFFICACY OF 5-ARYLAZO RHODANINES ADMINISTERED IN FEED FOR FOUR WEEKS IN THE CONTROL OF IMMATURE ASC'ARIDIA GALLI INFECTIONS Number Substituent on rhodanlne rlng Avg., Expt. A. gah'i Percent Number worm Etficacy, N0. ova per drug in rds number percent bird teed per lot at 4 3 position I B-pusltion weeks t G405. 650 Non-medlcated control r. 11.8 Methyl P-nitrophenylaz 0.10 10 0. 9 92. 4 do do.. 0. 05 ll) 1. 6 86. 4

8-121. 300 Non-medicated control 10 12.9

Methyl P-nitruphen az .05 1O 1. 3 89.9 do do 0. 025 10 3. 7 71. 3 Hydrogen d0 0. 05 10 1. 3 89. 9 do d 025 10 2.8 78.3

(1-209.. 300 Non-medicated control 11 39.2 Methyl a 12 1. l 97. 2 do. v 12 0.1 99.1 do 24'hloro, 4-nitr0pheny am... 0.05 12 0 100.0

The 5(P-nitrophenylazo) rhodanine compounds were tested under natural farm conditions and the results are tabulated in Table 5.

Table 5 rhodanines, 3-rnethyl, 5(P-nitrophenyla2/o) rhodanine was administered in a single oral capsule dose and results indicate that a single oral dose as high as 2900 mg./kg.

ACTIVITY OF GRADED LEVELS OF S-METiIYL, 5(P-NITROPHENYLAZO) RHO'DANINE AND RELATED HOMOLOGS ADMINISTERED IN FEED F OR TEN WEEKS AGAINST NATURAL INFECTIONS OF ASCARIDIA GALE! AT THE MOORMAN RESEARCH FARM Number Body Lb. Percent worm control at Farm 3-alkyl sub- Percent birds/lot 1 weight feed/lb. Point Weeks Expt. stituent drug in (110.) at surspread 1 N 0. feed 10 weeks viving Started Dead bird 4 6 8 l0 8l3B O 104 4 3. 73 2. 98 0. 10 104 2 3. 53 2. 59 94 57 97 98 100 0. 05 103 3 3. 70 2. 71 99 46 98 0. 25 2 3. 83 2. 52 131 95 94 98 100 0. 0125 104 4 3. 79 2. 61 11B 76 36 98 84613.-.. No drug 0 77 2 3. 66 2. 56

Methyl. O. 76 0 3. 80 2. 50 13B 78 98 98 98 d 0 76 1 3. 82 2. 51 131 40 6- 68 83 76 4 3. 80 2. I56 114 0 2O 25 47 76 2 3. 67 2. 71 96 69 74 B1 77 76 4 3. 84 2. 64 53 56 36 72 76 0 3. 69 2. 60 109 81 62 82 84 1 At two-Week intervals, 15 birds per lot were removed and killed for determination of worm burdens.

2 Point spread equals 100 body weight-feed efiicicncy).

EXAMPLE 4 Since one of the objectives of this invention is the conbody weight had no adverse elfect on survival of fiveweek-old pullets (Table 7). Therefore, the minimum 75 lethal dose is in excess of 2 900 trig/kg.

Table 6 BIRDS (EXPT. (Ir- 128) 5 Pereent drug in feed 0.05

Number birds per lot:

Initial Alive at 0 weeks Avg. body weight in 1b. at 9 weeks 1. Feed in lb. consumed per bird in 9 weeks 6. Lb. feed per lb. gain 3 Table 7 EFFEGI OF SINGLE ORAL CAPSULE ADMINISTRATION OF 3-METHYL, (P-NITROPHENYLAZO) RHODANINE ON MORTALITY AND WEIGHT GAINS IN FIVII-W'EEK- OLD \VHITE LEGHORN PULLETS (EXPT. G-128) COOCIDIO STAT EVALUATION OF of several different ways as illustrated by the following example:

EXAMPLE 5 A suitable basal broiler ration comprises Lbs. Ground yellow corn 67 Commercial chick concentrate (medicated) 33 Total 100 The commercial chick concentrate was a typical one containing soybean oil meal, alfalfa, meat scrap, fish meal, a complex mineral mix, vitamin sources and antibiotic supplement (penicillin) in usual known proportions to provide a high energy chick feed when mixed with corn. Into such a chick concentrate sufficient 5(nitrophenylazo) rhodanine is blended to give a concentration of from 0.0375% to 0.6% by weight.

The active ingredients may also be administered in the drinking water, in tablet form, or in capsules. However, incorporation of the active ingredients in the feed represents the preferred method.

In addition to exhibiting anthelmintic activity in poultry as set forth above, 3-n-butyl, 5(P-nitrophenylazo) rhodanine and 3-methy1, 5(2'methyl, 4nitrophenylazo) rhodanine also exhibit good coccidiostat activity. This is illustrated by the data contained in Table 8.

Table 8 5-ARYLAZO RHODANINES AGAINST MIXED INFECTIONS OF E.

TENELLA AND E. NEC'ATRIX Substltuent on rhodanine ring Number birds Number Percent Coccidio- Expt. Percent 000 sts per weight stat No. drug ird Dead 0! Percent gain in index 3-p0sitl0n 5-p0siti0n Initial coeciidiosurvival 7 days 040--.. 0 0 10 0 100 100 200 0 40, 000 10 7 7 23 0. 05 40, 000 10 7 30 21 9 0. 05 000 10 5 4 54 0. 05 40. 000 10 5 50 46 95 0. 05 40, 000 10 5 18 32 0. D5 40, 000 10 5 50 17 67 0. 05 40, 000 10 1 90 83 173 C 0 0 10 0 100 100 200 0 40, 000 10 5 50 -1 49 0. 005 40, 000 10 4 00 15 45 0. 01 40, 000 10 7 30 17 47 0.02 40, 000 10 1 58 148 0. 03 40, 000 10 1 90 60 150 0. 04 40, 000 10 0 82 182 0. 05 40, 000 10 0 100 58 158 0-90--.. 0 12 0 100 100 200 0 288,000 12 4 67 14 81 0. 05 P-nitrophenylam... 28B, 000 12 0 100 49 149 0. 025 do 288, 000 11 0 100 45 146 0. 05 2,4 dinltr0phenylaz0 288, 000 11 2 82 21 61 0. 05 2 chloro, 4 nitrophenylazm- 288, 000 12 1 92 3 89 0.05 2 methyl, 4 nitrophenylazm. %8. 000 12 0 100 42 142 0. 025 do 288. 000 12 1 92 22 114 0.0125 do-. ...-do 288, 000 11 2 82 19 101 The 5(nitrophenylazo) rhodanines, or mixtures thereof may be fed to poultry by including it in the ration in one 75 bed. be combined to provide an index 0! elfieaey called the Coccldiostat Index" which is day period iollowing infection. The maximum value of the EXAMPLE 6 Using the techniques for critical evaluation of anthelmintic activity described in Example 1, the activity of the 5 (nitrophenylazo rhodanines was studied in dogs naturally infected with hookworms and ascarids. Administration of the therapeutic dose in feed (approximately 100 mg./ kg.) over a 5-day period was used and the data presented in Table 9 illustrates the relative anthelrnintic activity of typical 5(nitrophenylazo) rhodanines. As seen in Table 10, administration of 3 methyl, 5(P-nitrophenylazo) rhodanine at low levels in feed for 15 days was effective in suppressing the production of ascarid and hookworm eggs. Thus, administration of these compounds administered continuously in the ration of dogs can prevent contamination of dog runs and thus break the life cycle of ascarids and hookworms in dogs in the same manner as described above in connection with poultry.

Table 9 about 0.2% by weight of a poultryration and from about 0.0088% to about 0.33% by weight of a dog ration:

as sehgm.

ANTIIELMINTIC ACTIVITY OF S-ARYLAZO RHODANINES ADMINISTERED IN FEED FOR FIVE DAYS AGAINST IIUOK'WORM AND ASCARID INFECTIONS IN NATU- RALLY INFECTED DOGS Substltuent on rliodantne rLng Total worms Total drug Parasite intake per Num- Eflicacy dog berdogs (percent) 3-poslti0n fi-positlon (mg/kg.) Initial autopsy Hookworm-.- Methyl P-nitrophenylazo 55. 2 402 0 100.0 90. 0 1 206 3 98. 100. 0 2 193 l 99. 5 110. 0 5 176 7 96. 0 d0 2 methyl, 4 nitrophenylazm. 110.0 1 4 0 100. 0

Ascaridd0 P-nitrophenylazo 9t}. 0 l 2 0 100. D 110.0 4 49 7 85. 7 do 107. 5 2 9 4 55. 6 o. 113,0 2 18 0 50.0 94. 5 l 6 1 83. 3 n-Butyl .do. 213. 0 l 7 0 100. 0 Methyl 2 methyl, 4 nitrophenylazm. 110. D 1 16 0 100. 0

Table 10 EFFECT OF 3-lVIETHYL 5(P-NITROPNENYLAZO) RHGDA NINE ADIMINISTERE AT LOW' LEVELS IN FEED FOR DAYS ON SHEDDING OF HOOKVVORM AND ASCARID EGGS (DOG EXPT. #6)

Drug intake Worm egg count (eggs per (mg/kg.) gram feces) Parasite Dog No. Start 13th day 15th day Per Total in trcat of treat postday 15 days merit ment treatment Ho0kw0rm 61B 8. 9 134. 2 225 0 0 62B 8. 5 127. 4 420 0 0 63B 7. 8 117.3 645 0 64B 7. 4 1 ll. 5 255 0 0 65B 6. 7 99. 9 435 0 0 66B 6. 6 98 3 0 67B 4. 4 66. 4 780 D 0 Ascarld 61B 8. 9 134. 2 45 0 0 62B 8. 5 127. 4 945 0 45 67B 4. 4 66. 4 150 270 240 X No sample.

The 5 (nitrophenylazo) rhodanine compounds useful in the present invention and methods of preparing the same, are disciosed in Patent No. 2,952,673. This present application is a continuation-in-part of prior application Serial No. 60,293, filed October 4, 1960, and of Serial No. 799,417, filed March 16, 1959, both of which are now abandoned.

In view of the foregoing disclosure, those skilled in the art will be able to practice the invention either as disclosed or in one or more other suitable and obvious manners.

I claim:

1. A medicated feed composition for treatment, control and prevention of ascarids in poultry and dogs, and hook- Worms in dogs comprising alimentary material containing sufiicient 5(nitr0phenylazo) rhodanine of the following structural formula to provide from about 0.00625% to in which the compound is 3ethyl, 5(P-nitrophenylazo) rhodanine.

4. A medicated feed composition as defined in claim 1 in which the compound is 3-n-butyl 5(P-nitrophenylazo) rhodanine.

5. A medicated feed composition as defined in claim 1 in which the compound is 3-methyi, 5(2' methyl, 4' nitrophenylazo) rhodanine.

6. A medicated feed composition as defined in claim I in which the compound is 3-methyl, 5(2' chioro, 4' nitrophenylazo) rhodanine.

7. Method of treating, controlling and preventing ascarids in poultry and dogs, and hookworms in dogs which comprises feeding poultry a ration containing from about 0.00625% to about 0.2% by weight and dogs a ration containing from about 0.0088% to about 0.33% by weight of 5 (nitrophenylazo) rhodanine of the following structural formula:

in which R represents a member selected from the group consisting of hydrogen, methyl, ethyl, propyl, and butyl radicals, and X represents a member selected from the group consisting of methyl, chloro and nitro substituents.

8. The method of treatment, control and prevention of ascarids in poultry and dogs, and hookworms in dogs as defined in claim 7 in which the compound is B-methyl, 5 (P-nitrophenylazo) rhodanine.

9. The method of treatment, control and prevention of ascarids in poultry and dogs, and hookworms in dogs as defined in claim 7 in which the compound is 3-ethyl, 5(1 nitrophenylazo) rhodanine.

10. The method of treatment, control and prevention of ascarids in poultry and dogs, and hookworms in dogs 1 1 1 2 as defined in claim 7 in which the compound is 3-n-buty1, ration containing from about 0.02% to about 0.2% by 5(P-nitropheny1azo) rhodanine. weight of n-butyl 5(P-nitrophenylazo) rhodanine and 3- 11. The method of treatment, control and prevention methyl 5(2methyl, 4'nitrophenylazo) rhiodanine. of ascarids in poultry and dogs, and hookworrns in dogs as defined in claim 7 in which the compound is 3-rnethy1, 5 References Cited in the file of this patent 5 (2' methyl, 4' nitrophenylazo) rhodanine. UNITED STATES PATENTS 12. The method of treatment, control and prevention of ascarids in poultry and dogs, and hookworms in dogs 2743211 Bashour 1956 as defined in claim 7 in which the compound is S-rnethyl, 5(2' chloro, 4 nitrophenylazo) rhodanine. 10 OTHER REFERENCES Mackie, Brit. J. Pharm. 8; Chemotherapy, vol. 10, pp.

13. The method ftre tin controll'n and r ventin o a g lg N g 19,1955.

coccidiosis in poultry which comprises feeding poultry a. 

1. A MEDICATED FEED COMPOSITION FOR TREATMENT, CONTROL AND PREVENTION OF ASCARIDS IN POULTRY AND DOGS, AND HOOKWORMS IN DOGS COMPRISING ALIMENTARY MATERIAL CONTAINING SUFFICIENT 5 (NITROPHENYLAZO) RHODANINE OF THE FOLLOWING STRUCTURAL FORMULA TO PROVIDE FROM ABOUT 0.00625% TO ABOUT 0.2% BY WEIGHT OF A POULTRY RATION AND FROM ABOUT 0.0088% TO ABOUT 0.33% BY WEIGHT OF A DOG RATION: 